Indian Journal of Geo-Marine Science Vol.45(1), January 2016,pp. 161-166

First record and colour banding pattern of karachiensis (Winckworth, 1930) (: : ) from Gujarat coast, India

Paresh Poriya, Bhavik Vakani & Rahul Kundu* Department of Biosciences, Saurashtra University, Rajkot-360005, Gujarat, India [E-mail: [email protected]]

Received 26 March 2014; revised 10 October 2014

Present communication reports the first record of a species Cellana karachiensis (Winckworth, 1930) from Western Indian coast which was previously identified as (Born, 1778). The species was initially presumed as Cellana radiata as reported earlier by many authors. The results of the DNA barcoding pointed out that it is 99 % identical with Cellana karachiensis, and not C. radiata. C. karachiensis is till date reported only from Gulf of Oman and thus, this study reports the first record of Cellana karachiensis (Mollusca: Nacellidae) from Veraval coast, Gujarat State, India. Three different color banding patterns were observed in this . There was a little variation found in DNA barcoding as Type X match up to 99.69 %, Type Y and Type Z match up to 99.54 % with previously submitted DNA sequences of C. Karachiensis. We initially proposing these as three distinct subtypes (X, Y and Z) of the Cellana and species karachiensis, till further evidences arises as being same or different species.

[Keywords: Cellana karachiensis, first record, molecular evidence, DNA Barcoding, Western Indian coast]

Introduction Cellana karachiensis, no previous records In India, about 5070 species of molluscs were found for its occurrence in Indian coastal reported amongst the world molluscan faunal areas. These from the Kathiawar coast resources which contains 66535 identified was previously identified as Cellana radiata molluscan species1. In India, a total of 3271 based on its external shell morphology. But, species of marine molluscs, belonging to 220 recently Nakano & Ozawa have done the DNA families and 591 genera were identified. This barcode of Limpets and their phylogeography includes around 1900 gastropods, 1100 of the order Patellogastropoda worldwide and bivalves, 210 cephalopods, 41 in this report C. karachiensis found distributed polyplacophores and 20 are scaphopods. only in Gulf of Oman13. Amongst them, Patellogastropoda (limpets) represent a diverse group and shows a Ecology of Cellana radiata and the colour worldwide distribution restricted exclusively banding patterns on its shell was magnificently to marine habitats. Members of described earlier on Kathiawar coast14,15,16,17,18. Patellogastropoda are currently classified in There are three colour banding patterns were five families, viz. , Nacellidae, reported earlier on shell of C. radiata. Present , and . Limpets study was undertaken to check whether these are mostly found in intertidal zone throughout patterns are of the same species or they are the world oceans from tropical to Polar different subspecies of the same genus as they Regions and play an important role in resembles very close external morphological intertidal marine ecosystems2,3. characters. This study revolves around the hypothesis that whether the having External shell morphology have been three different colour banding patterns are traditionally used in of limpet same species, different subspecies of same species but sometimes variation in shell’s genus, or, they are different species altogether. shape and size leads to incorrect In order to test this hypothesis, animals of identification4,5,6,7,8,9. But in recent times, different size groups having different colour molecular characters like DNA sequence data banding patterns were collected from an have been widely used as an important tool for undisturbed habitat of Veraval coast (India) proper identification10,11,12. In case of a limpet, off Arabian Sea and identified. For 162 INDIAN J. MAR. SCI. VOL. 45, NO.1 JANUARY 2016 confirmation of exact species identification communication, we are reporting it from DNA barcoding technique was applied using Veraval, Gujarat State, India. As per the IUCN mitochondrial cytochrome c oxidase subunit I Status, this species has not assessed for the gene (COI) and identification of this species IUCN Red List so far, but is in the ‘Catalogue was done by matching the DNA sequence with of Life’21. Regarding the habitat and external the earlier submitted DNA sequence in BOLD morphology, C. Karachiensis found abundant (Barcoding of Life Database) System and in the spray zone and comparatively low in GenBank. upper littoral zone, while rare in middle and lower littoral zones. This animal is very Materials and Methods sluggish in nature and generally found settled on the walls of the pools and the crevices. Genus Cellana is widely distributed These animals are also present in various kinds throughout rocky intertidal zones of Indian of micro-habitats like crevices, gullies, rock coast. In the present study, samples of C. pools, but sparse on rocks encrusted with radiata having different banding patterns, and seaweeds. Shell measurement termed as Types X, Y and Z, were collected recorded was average length of 36mm, breadth from the intertidal zone of Veraval (210 35’ N, 30mm, height 12mm, weight 225mg, distance 690 36’ E) on the Kathiawar Peninsular from apex to anterior margin 18mm and coastline off Arabian Sea. Collected samples distance from apex to posterior margin 24mm. were preserved in 99% analytical grade Shell of C. Karachiensis shows three different ethanol and sent to the Rajiv Gandhi Centre color patterns (Fig. 1). The ‘X’ type shows for Biotechnology, Kerala (Government of alternate black and white broad bands towards India, Ministry of Science & Technology, the peripheral region of shell opening. The Department of Biotechnology) for DNA umbo of this type of shell is slightly yellowish barcoding. DNA Barcoding was done by using with somewhat blunt end. Second ‘Y’ type universal primer COX119,20. New sequence shows alternate black and white very narrow obtained in this study was compared in BOLD bands or thin lines/ridges on the entire surface System and GenBank where it was aligned of the shell. The black bands sometimes show with publically available and previously irregular distribution. The umbo in this case is confirmed sequences of different animal pointed. The third ‘Z’ type is characterized by species. distinct elevated white ridges on the shell. These ridges are not continuous but interrupted at several places by black Results and Discussions ridges/lines. These white ridges sometimes are accompanied by black spots near their basal This study demonstrates the utility of DNA portions. markers besides the morphologic characteristics in animal taxonomy and As far as the DNA barcoding is concerned, systematics. The collected species, previously there was little variation found as Type-X was identified as C. radiata is now re-identified as matching up to 99.69 % and Type-Y and Cellana karachiensis after the submitting Type-Z up to 99.54 % with previously obtained DNA sequences in BOLD System submitted DNA sequences of C. Karachiensis and GenBank. C. karachiensis is a species of (Table-4, Table-5, and Table-6). DNA marine gastropod mollusc of family Nacellidae sequence of Type – B and Type – C showed and commonly known as the moon snail variation next to C. Karachiensis, while, it was (Table-1, Table-2, Table-3). Present comparing with earlier submitted top 20 investigation provides new account of a limpet matches of gene sequences of nearby species. species Cellana karachiensis for Indian coast. This little genetic variations also generates This also indicating the necessity of re- doubts that weather the animals having three evaluating different species identification of different colour banding patterns are same various fauna by the molecular characteristics species, different subspecies of same genus or using DNA barcoding techniques. they are different species altogether. It was also interesting to note that these three types C. Karachiensis was reported only from Oman shell colour banding patterns seemed to be (Western Indian Ocean)13, and in this distinct only after the achievement of PORIYA et al.: FIRST RECORD OF CELLANA KARACHIENSIS FROM INDIAN COAST 163

particular body size. Spats and smaller size the shell surface with irregular distribution of individuals only show distinct white ridges on small black spots or lines.

Table-1. Taxonomic Details from BOLD System for karachiensis till further evidences emerges as Type-X upon submission of DNA sequence. their being same or different species.

Taxonomic Probability of Taxon Assignment Figure-1. Different colour banding patterns in the shells Level Placement (%) of Cellana karachiensis. Phylum Mollusca 100 Class 100 Order Docoglossa 100 Family Nacellidae 100 Genus Cellana 100 Species Cellana karachiensis 99.69

Table-2. Taxonomic Details from BOLD System for Type-Y upon submission of DNA sequence.

Taxonomic Probability of Taxon Assignment Level Placement (%) Phylum Mollusca 100 Class Gastropoda 100 Order Docoglossa 100 Family Nacellidae 100 Genus Cellana 100 Species Cellana karachiensis 99.54

Table-3. Taxonomic Details from BOLD System for Type-Z upon submission of DNA sequence.

Taxonomic Probability of Taxon Assignment Level Placement (%) Phylum Mollusca 100 Class Gastropoda 100 Order Docoglossa 100 Family Nacellidae 100 Genus Cellana 100 Species Cellana karachiensis 99.54

Population composition of ‘X’, ‘Y’ and ‘Z’ banding patterns of C. karachiensis were also found varied and showed season wise definite 14 pattern . Creese and Underwood have concluded that shell-banding patterns in the trochid gastropod Austrocochles constricta are Acknowledgment 22 primarily governed by environmental factors . Authors are thankful to UGC, Govt. of India, On the other hand, the banding pattern of New Delhi, for supporting this study through pigmentation in some intertidal pulmonates its Centre of Advanced Study Programme has been shown to be due to a genetic 23,24 (CAS Phase-I) and for the Meritorious polymorphism while Innes and Haley Research Fellowship awarded to the senior explain it as inheritance of a shell-color 25 author. Authors are also thankful to Rajiv polymorphism in the mussel . We thus, Gandhi Centre for Biotechnology- Kerala initially proposing these as three distinct types (Govt. of India) for providing DNA barcoding (X, Y and Z) of the genus Cellana and species services. 164 INDIAN J. MAR. SCI. VOL. 45, NO.1 JANUARY 2016

Table-4. Similarity scores of Top 20 Matches as per BOLD System for Type-X upon submission of DNA sequence.

Phylum Class Order Family Genus Species Similarity (%) Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 99.69 Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 99.08 Mollusca Gastropoda Docoglossa Nacellidae 90.32 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.94 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.93 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.82 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.79 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.79 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.78 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.63 Mollusca Gastropoda Docoglossa Nacellidae Cellana taitensis 88.63 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.48 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Lepetidae fulva 87.25

Table-5. Similarity scores of Top 20 Matches as per BOLD System for Type-Y upon submission of DNA sequence.

Phylum Class Order Family Genus Species Similarity (%) Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 99.54 Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 98.92 Mollusca Gastropoda Docoglossa Nacellidae Cellana eucosmia 90.17 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.09 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.09 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.93 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.82 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.79 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.78 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.63 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.63 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.48 Mollusca Gastropoda Docoglossa Nacellidae Cellana taitensis 88.48 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.33 Mollusca Gastropoda Archaeogastropoda Lepetidae Iothia fulva 87.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.25 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.1

PORIYA et al.: FIRST RECORD OF CELLANA KARACHIENSIS FROM INDIAN COAST 165

Table-6. Similarity scores of Top 20 Matches as per BOLD System for Type-Z upon submission of DNA sequence.

Phylum Class Order Family Genus Species Similarity (%) Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 99.54 Mollusca Gastropoda Docoglossa Nacellidae Cellana karachiensis 98.92 Mollusca Gastropoda Docoglossa Nacellidae Cellana eucosmia 90.48 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 89.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89.17 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89.11 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89.09 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 89.02 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.94 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.94 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata capensis 88.94 Mollusca Gastropoda Docoglossa Nacellidae Cellana taitensis 88.79 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.63 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 88.48 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.56 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.4 Mollusca Gastropoda Docoglossa Nacellidae Cellana radiata 87.4 Mollusca Gastropoda Archaeogastropoda Lepetidae Iothia fulva 87.4

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